Journal
MOLECULAR AND CELLULAR BIOCHEMISTRY
Volume 247, Issue 1-2, Pages 171-176Publisher
KLUWER ACADEMIC PUBL
DOI: 10.1023/A:1024167731074
Keywords
ascorbic acid; ascorbate 2-phosphate; alpha-tocopherol; H4IIE cells; cis-parinaric acid; lipid peroxidation
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Funding
- NIA NIH HHS [AG 16236] Funding Source: Medline
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Ascorbic acid, or vitamin C, can recycle alpha-tocopherol in lipid bilayers, but even sparing of alpha-tocopherol has not been a consistent finding in intact cells. Therefore, we tested the ability of ascorbate loading to spare alpha-tocopherol and to prevent lipid peroxidation of cultured H4IIE rat liver cells. Although alpha-tocopherol was undetectable in H4IIE cells, its cell content was increased by overnight incubation with alpha-tocopherol in culture. Cells incubated with ascorbate 2-phosphate accumulated ascorbate to concentrations as high as 0.6 mM after overnight loading, but also released ascorbate into the medium. Ascorbate loading of alpha-tocopherol-treated cells spared alpha-tocopherol in a concentration-dependent manner during overnight incubation. Lipid peroxidative damage, measured as a decrease in fluorescence of cell-bound cis-parinaric acid, was decreased in cells loaded with either alpha-tocopherol or ascorbate 2-phosphate, and showed an additive effect. These results suggest that ascorbate loading of H4IIE cells spares cellular alpha-tocopherol and either directly or through recycling of alpha-tocopherol prevents lipid peroxidative damage due to oxidant stress in culture.
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